Twist1对胶质瘤样干细胞的增殖和侵袭影响

The effect of Twist1 on proliferation and invasion of glioma stem-like cells

目的研究Twist1对胶质瘤样干细胞增殖和侵袭影响。方法通过无血清干细胞培养基方法从原代培养的多形性胶质母细胞瘤中获得悬浮生长的肿瘤细胞球,利用免疫荧光方法及血清诱导分化方法对培养的肿瘤球进行鉴定;通过Western blot方法比较胶质瘤样干细胞和原代培养的多形性胶质母细胞瘤中Twist1表达;SiRNA抑制Twist1表达后,应用CCK-8方法及Transwell方法检测Twist1对胶质瘤样干细胞增殖和侵袭影响。结果无血清干细胞培养基方法可以从原代培养的多形性胶质母细胞瘤中诱导出悬浮生长的肿瘤细胞球;肿瘤细胞球表达肿瘤干细胞标记物CD133和Sox2,在含血清培养基中可以分化为表达神经元和神经胶质细胞标记物的细胞;胶质瘤样干细胞高度表达Twist1;利用siRNA抑制Twist1后,可以抑制胶质瘤样干细胞增殖和侵袭。结论Twist1可能是维持胶质瘤样干细胞干性的关键基因。

Objective To study the effect of Twistl on proliferation and invasion of glioma-like stem cells. Methods The tumor cell spheres with suspended growth were obtained from primary cultured glioma ceils by serum-free stem cell culture medium. Immunofluorescence and serum induced differentiation were used to identify the tumor cell spheres. The expression of Twistl protein was determined by Western blot. The proliferation and invasion of glioma-like stem cells were detected by CCK-8 method and Transwell method. Results Serum-free stem cell culture medium method could induce tumor cell spheres from primary cultured glioma cells. The cell spheres were expressed CD133 and Sox2, and differentiated into cells expressing neurons and glial cell markers in serum-containing medium. Glioma stem-like cells were highly expressed Twistl. Inhibition of protein expression of Twistl by siRNA transfection, the proliferation and invasion of glioma stem-like cells were inhibited. Conclusion Twistl may be the key gene for the maintenance of glioma stem-like cells and can promote their proliferation and invasion.